title: Tracking a refined eIF4E-binding motif reveals Angel1 as a new partner of eIF4E.
abstract: The initiation factor 4E (eIF4E) is implicated in most of the crucial steps of the mRNA life cycle and is recognized as a pivotal protein in gene regulation. Many of these roles are mediated by its interaction with specific proteins generally known as eIF4E-interacting partners (4E-IPs), such as eIF4G and 4E-BP. To screen for new 4E-IPs, we developed a novel approach based on structural, in silico and biochemical analyses. We identified the protein Angel1, a member of the CCR4 deadenylase family. Immunoprecipitation experiments provided evidence that Angel1 is able to interact in vitro and in vivo with eIF4E. Point mutation variants of Angel1 demonstrated that the interaction of Angel1 with eIF4E is mediated through a consensus eIF4E-binding motif. Immunofluorescence and cell fractionation experiments showed that Angel1 is confined to the endoplasmic reticulum and Golgi apparatus, where it partially co-localizes with eIF4E and eIF4G, but not with 4E-BP. Furthermore, manipulating Angel1 levels in living cells had no effect on global translation rates, suggesting that the protein has a more specific function. Taken together, our results illustrate that we developed a powerful method for identifying new eIF4E partners and open new perspectives for understanding eIF4E-specific regulation.
authors.affiliation: UPMC Univ Paris 06, UMR 7150, Mer et Santé, Station Biologique, F-29680 Roscoff, France, CNRS, UMR 7150, Mer et Santé, Station Biologique, F-29680 Roscoff, France. Université Européenne de Bretagne, Bretagne, Roscoff, France, INSERM, UMR 1037, Centre de Recherche en Cancérologie de Toulouse, Toulouse 31432, France, UPMC Univ Paris 06, UMR 7139, Végétaux Marins et Biomolécules, Station Biologique, F-29680 Roscoff, France, CNRS, UMR 7139, Végétaux Marins et Biomolécules, Station Biologique, F-29680 Roscoff, France, UPMC Univ Paris 06, FR2424, ABiMS, Station Biologique, F-29680 Roscoff, France and CNRS, FR2424, ABiMS, Station Biologique, F-29680 Roscoff, France.
authors.forename: Pauline
authors.initials: P
authors.lastname: Gosselin
authors.forename: Yvan
authors.initials: Y
authors.lastname: Martineau
authors.forename: Julia
authors.initials: J
authors.lastname: Morales
authors.forename: Mirjam
authors.initials: M
authors.lastname: Czjzek
authors.forename: Virginie
authors.initials: V
authors.lastname: Glippa
authors.forename: Isabelle
authors.initials: I
authors.lastname: Gauffeny
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Morin
authors.forename: Gildas
authors.initials: G
authors.lastname: Le Corguillé
authors.forename: Stephane
authors.initials: S
authors.lastname: Pyronnet
authors.forename: Patrick
authors.initials: P
authors.lastname: Cormier
authors.forename: Bertrand
authors.initials: B
authors.lastname: Cosson
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000145:classification / Q000378:metabolism*; D003593:Cytoplasm / Q000737:chemistry; D004721:Endoplasmic Reticulum / Q000737:chemistry; D039561:Eukaryotic Initiation Factor-4E / Q000032:analysis / Q000378:metabolism*; D006056:Golgi Apparatus / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D054730:Protein Interaction Domains and Motifs; D012260:Ribonucleases / Q000145:classification
chemical_list: C584138:Angel1 protein, human; D002352:Carrier Proteins; D039561:Eukaryotic Initiation Factor-4E; D012260:Ribonucleases; C445084:mRNA deadenylase